Anti-6X His tag® antibody (ab9108)
Key features and details
- Rabbit polyclonal to 6X His tag®
- Suitable for: WB, ICC/IF, ELISA, ChIP
- Reacts with: Species independent
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
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- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-6X His tag® antibody
See all 6X His tag® primary antibodies -
Description
Rabbit polyclonal to 6X His tag® -
Host species
Rabbit -
Specificity
ELISA: Antibody specificity was verified by ELISA against peptide conjugated to BSA (HHHHHH-BSA). A 1:25,000 dilution of the antibody gave an OD=1.0 in a 15 minute reaction using HRP-conjugated Goat anti-Rabbit IgG at 1:20,000 and TMB as the substrate. Appropriate specificity controls were run. -
Tested applications
Suitable for: WB, ICC/IF, ELISA, ChIPmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
Synthetic peptide corresponding to 6X His tag® conjugated to keyhole limpet haemocyanin.
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Positive control
- ICC/IF: CHO cells transfected with 12 tags constructs (CHO-12 tags) and HEK293 expressing His tag. WB: HeLa transfected with His-tagged gene, Epitope Tag Positive Control whole cell lysates, purified 12-Tag and cell lysates of CHO-12 Tags.
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General notes
This antibody recognizes both C-terminal and N-terminal tags.
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).
See other anti-rabbit secondary antibodies that can be used with this antibody.
HIS-TAG® is a trademark of EMD Biosciences, Inc.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.1% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab9108 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (14) |
Use a concentration of 1 µg/ml.
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| ICC/IF | (2) |
1/500 - 1/1000.
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| ELISA | (2) |
1/1000.
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| ChIP |
Use at an assay dependent concentration. ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads.
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| Notes |
|---|
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WB
Use a concentration of 1 µg/ml. |
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ICC/IF
1/500 - 1/1000. |
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ELISA
1/1000. |
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ChIP
Use at an assay dependent concentration. ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads. |
Target
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Relevance
The H-H-H-H-H-H motif is used as a tag on many recombinant proteins to facilitate purification. His-tags can be fused to the amino- or carboxy- termini of proteins in transfected or transformed cells. -
Cellular localization
Depends upon the localization of the parent protein tagged with hexahistidine. -
Alternative names
- 6 His epitope tag antibody
- Hexa His tag antibody
- HHHHHH epitope tag antibody
see all
Images
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Western blot - Anti-6X His tag® antibody (ab9108)All lanes : Anti-6X His tag® antibody (ab9108) at 1 µg/ml
Lane 1 : Purified 12-Tag at 2 µg
Lane 2 : Purified 12-Tag at 0.2 µg
Lane 3 : The cell lysates of CHO-12 Tags at 2 µg
Lane 4 : The cell lysates of CHO at 2 µg
Secondary
All lanes : Goat anti-rabbit, HRP conjugated at 0.2 µg/mlLane 1 and 4: Molecular weight Standards.
Primary and secondary antibody incubation at RT for 1 hour.
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Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody (ab9108)Immunofluorescent analysis of 4% paraformaldehyde (PFA) -fixed, permeabilized with 0.1% Triton X-100 in CHO cells transfected with 12 tags constructs (CHO-12 tags) labelling 6xHIS with ab9108 at 1/1000 dilution (1µg/mL), followed by Donkey Anti-rabbit IgG (H&L), Alexa Fluor® 594 antibody at 1/500 dilution (4 µg/mL) (Red). Nucleus was counterstained with DAPI (Blue). Bottom panel images showed Parallel staining in parental CHO cell line.
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Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody (ab9108)This image is courtesy of Vladimir Milenkovicab9108 staining CACNB3 with His tag expressed in HEK293 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/500 in 1% goat serum, 0.1% TX100 and PBS) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody (1/750).
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ChIP - Anti-6X His tag® antibody (ab9108)A stably transfected 293T human cell line harbouring the GAL4 upstream activation sequence was transiently transfected with a Myc or His - tagged GAL4 DNA Binding Domain construct. 48 hours post transfection Chromatin was prepared according to the Abcam X-ChIP protocol. The ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads. A non-specific antibody was used as the negative control. The immunoprecipitated DNA was quantified by real time PCR (SYBR Green approach). -
Western blot - Anti-6X His tag® antibody (ab9108)This image is courtesy of an anonymous AbreviewAll lanes : Anti-6X His tag® antibody (ab9108) at 1/4000 dilution
Lane 1 : HeLa whole cell lysate - transfected with empty vector
Lane 2 : HeLa whole cell lysate - transfected with His-tagged gene
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 35 kDa why is the actual band size different from the predicted?
Exposure time: 30 seconds
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Western blot - Anti-6X His tag® antibody (ab9108)Use of ab9108 in western blotting at 1:1000 against Epitope Tag Positive Control lysate (ab4524).
Lane 1: 2 µl lysate
Lane 2: 1 µl lysate
Lane 3: 0.5 µl lysate
Lane 4: 0.25 µl lysate Use of ab9108 in western blotting at 1:1000 against E.coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 11 different epitope tag (ab5395). Lane 1: 2 µl lysate Lane 2: 1 µl lysate Lane 3: 0.5 µl lysate Lane 4: 0.25 µl lysate
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (150)
ab9108 has been referenced in 150 publications.
- Bordet G et al. Poly(ADP-ribosyl)ating pathway regulates development from stem cell niche to longevity control. Life Sci Alliance 5:N/A (2022). PubMed: 34949666
- Meng F et al. Lysosomal iron recycling in mouse macrophages is dependent upon both LcytB and Steap3 reductases. Blood Adv 6:1692-1707 (2022). PubMed: 34982827
- Ercanoglu MS et al. No substantial preexisting B cell immunity against SARS-CoV-2 in healthy adults. iScience 25:103951 (2022). PubMed: 35224466
- Martinez B et al. Pervasive Transcription-coupled DNA repair in E. coli. Nat Commun 13:1702 (2022). PubMed: 35354807
- Azulay G et al. A dual-function phage regulator controls the response of cohabiting phage elements via regulation of the bacterial SOS response. Cell Rep 39:110723 (2022). PubMed: 35443160